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用于胸膜固定术的不同硬化剂的炎症反应评估中的原代人胸膜细胞培养。

Primary human mesothelial cell culture in the evaluation of the inflammatory response to different sclerosing agents used for pleurodesis.

机构信息

Department of Internal Medicine, Pulmonary Diseases & Allergy, Medical University of Warsaw, Warsaw, Poland.

出版信息

Physiol Rep. 2021 Apr;9(8):e14846. doi: 10.14814/phy2.14846.

Abstract

The mechanisms of chemical pleurodesis are still not fully explained. We aimed to evaluate the feasibility of using primary biopsy-derived human mesothelial cells to establish an in vitro culture and to assess the response of pleural mesothelial cells to different sclerosing agents. Talc, povidone-iodine, doxycycline, and TGF-β were used at different doses to stimulate pleural mesothelial cells. After 6 and 24 h, mRNA expression of interleukin (IL)-1β, IL-6, IL-8, TGF-β, MCP-1, IL-17A, and MMP9 was measured in cultured cells, and the protein level of IL-1β, IL-6, and IL-8 was measured in the culture supernatant. The most pronounced response was observed after talc exposure. It was expressed as an increase in IL-1β concentration in culture supernatant after 24 h of higher talc dose stimulation compared to 6 h of stimulation (17.14 pg/ml [11.96-33.32 pg/ml] vs. 1.84 pg/ml [1.81-1.90 pg/ml], p = 0.02). We showed that culture pleural mesothelial cells isolated from pleura biopsy specimens is feasible. Inflammatory responses of mesothelial cells to different sclerosants were highly variable with no consistent pattern of mesothelium reaction neither in terms of different sclerosing agents nor in the time of the most significant reaction. We demonstrated that pro-inflammatory mesothelial response includes an increase in IL-1β mRNA expression and protein production. This may suggest the role of IL-1β in the formation and maintenance of the inflammatory response during pleurodesis.

摘要

化学性胸膜固定术的机制尚不完全清楚。本研究旨在评估使用原发性活检衍生的人胸膜间皮细胞建立体外培养的可行性,并评估胸膜间皮细胞对不同硬化剂的反应。滑石粉、聚维酮碘、强力霉素和 TGF-β 以不同剂量刺激胸膜间皮细胞。6 和 24 h 后,测量培养细胞中白细胞介素 (IL)-1β、IL-6、IL-8、TGF-β、MCP-1、IL-17A 和 MMP9 的 mRNA 表达,并测量培养上清液中 IL-1β、IL-6 和 IL-8 的蛋白水平。在滑石粉暴露后观察到最明显的反应。表现为与 6 h 刺激相比,高剂量滑石粉刺激 24 h 后培养上清液中 IL-1β 浓度增加(17.14 pg/ml [11.96-33.32 pg/ml] 比 1.84 pg/ml [1.81-1.90 pg/ml],p=0.02)。我们表明,从胸膜活检标本分离的培养胸膜间皮细胞是可行的。不同硬化剂间皮细胞的炎症反应差异很大,无论是不同的硬化剂还是反应最明显的时间,间皮细胞反应均无一致模式。我们证明,促炎间皮反应包括 IL-1β mRNA 表达和蛋白产生增加。这可能表明 IL-1β 在胸膜固定术中炎症反应的形成和维持中起作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c15/8087983/9f374b15a6e9/PHY2-9-e14846-g002.jpg

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